The invention relates to the area of tumor suppression. More particularly, the invention relates to tumor suppressor genes and proteins.
Mutations in tumor suppressor genes play an important role in the development of neoplasias. Manipulation of tumor suppressor gene expression can be used to prevent or treat neoplasias. Detection of mutations in tumor suppressor genes can also be used to detect neoplastic cells and genetic predispositions to neoplasias. Thus, there is a need in the art for the identification of mammalian tumor suppressor genes which can be used to develop methods of diagnosing, prognosing, and treating neoplastic cells in humans and other mammals.
It is an object of the invention to provide an isolated and purified deep orange protein.
It is another object of the invention to provide an isolated and purified deep orange polypeptide.
It is yet another object of the invention to provide a deep orange fusion protein.
It is still another object of the invention to provide a preparation of antibodies which specifically bind to a deep orange protein.
It is yet another object of the invention to provide an isolated and purified subgenomic polynucleotide.
It is still another object of the invention to provide an expression construct for expressing all or a portion of a deep orange protein.
It is even another object of the invention to provide a mammal which does not express an endogenous wild-type deep orange gene in one or more tissues.
It is another object of the invention to provide a method to aid in the diagnosis or prognosis of cancer in a mammal.
It is yet another object of the invention to provide a method to aid in detecting a genetic predisposition to neoplasia in a mammal.
It is still another object of the invention to provide a method of identifying a human chromosome 15.
It is even another object of the invention to provide a method of identifying a mouse chromosome 2.
These and other objects of the invention are provided by one or more of the embodiments described below.
One embodiment of the invention provides an isolated and purified deep orange protein. The deep orange protein has an amino acid sequence which is at least 95% identical to the amino acid sequence shown in SEQ ID NO:4.
Another embodiment of the invention provides an isolated and purified deep orange polypeptide. The deep orange polypeptide comprises at least eight contiguous amino acids. The amino acid sequence of the at least eight contiguous amino acids is at least 95% identical to the amino acid sequence shown in SEQ ID NO:4.
Yet another embodiment of the invention provides a deep orange fusion protein. The fusion protein comprises a first protein segment and a second protein segment fused together by means of a peptide bond. The first protein segment consists of at least eight contiguous amino acids of a deep orange protein. The amino acid sequence of the deep orange protein is at least 95% identical to the amino acid sequence shown in SEQ ID NO:4.
Still another embodiment of the invention provides a preparation of antibodies which specifically bind to a deep orange protein. The amino acid sequence of the deep orange protein is at least 95% identical to the amino acid sequence shown in SEQ ID NO:4.
Even another embodiment of the invention provides an isolated and purified subgenomic polynucleotide. The polynucleotide comprises at least 1340 contiguous nucleotides and encodes an amino acid sequence which is at least 95% identical to the amino acid sequence shown in SEQ ID NO:4.
Yet another embodiment of the invention provides an expression construct for expressing all or a portion of a deep orange protein. The expression construct comprises a promoter and a polynucleotide segment. The promoter is not one which initiates transcription of a deep orange gene in normal human cells. The polynucleotide segment is located downstream from the promoter and encodes at least eight contiguous amino acids of a deep orange protein. The deep orange protein has at least 95% identity with the amino acid sequence shown in SEQ ID NO:4. Transcription of the polynucleotide segment initiates at the promoter.
Still another embodiment of the invention provides a mammal which does not express an endogenous wild-type deep orange gene in one or more tissues. The tissues are selected from the group consisting of: spleen, thymus, prostate, testis, ovary, small intestine, mucosal lining of the colon, heart, brain, placenta, liver, skeletal muscle, pancreas, lymph node, appendix, peripheral blood leukocytes, and bone marrow.
Even another embodiment of the invention provides a method to aid in the diagnosis or prognosis of cancer in a mammal. The method comprises comparing a deep orange gene, mRNA, or protein in a first tissue of a mammal suspected of being neoplastic with a deep orange gene, mRNA, or protein in a second tissue of the mammal which is normal. A difference between the deep orange gene, mRNA, or protein in the first and second tissues indicates a mutation in the deep orange gene in the first tissue of the mammal.
Another embodiment of the invention provides a method to aid in detecting a genetic predisposition to neoplasia in a mammal. The method comprises comparing a deep orange gene, mRNA, or protein in a fetal tissue of a mammal with a wild-type deep orange gene, mRNA, or protein. A difference between the deep orange gene, mRNA, or protein in the fetal tissue of the mammal and the wild-type deep orange gene, mRNA, or protein indicates a genetic predisposition to neoplasia in the mammal.
Yet another embodiment of the invention provides a method of identifying a human chromosome 15. The method comprises the steps of contacting a preparation of metaphase human chromosomes with a nucleotide probe and detecting a chromosome which specifically hybridizes to the nucleotide probe. The probe comprises at least 12 contiguous nucleotides selected from the nucleotide sequence shown in SEQ ID NO:1. A chromosome which specifically hybridizes to the nucleotide probe is identified as a human chromosome 15.
Even another embodiment of the invention provides a method of identifying a mouse chromosome 2. The method comprises the steps of contacting a preparation of metaphase mouse chromosomes with a nucleotide probe and detecting a chromosome which specifically hybridizes to the nucleotide probe. The probe comprises at least 12 contiguous nucleotides selected from the nucleotide sequence shown in SEQ ID NO:3. A chromosome which specifically hybridizes to the nucleotide probe is identified as a mouse chromosome 2.
The present invention thus provides the art with novel mammalian tumor suppressor genes, called xe2x80x9cdeep orange.xe2x80x9d Mammalian deep orange genes and proteins can be used, inter alia in methods to aid in diagnosing and prognosing neoplasia, to detect predispositions to neoplasia, to create animal models which can be used to develop therapeutic treatments for neoplasia, and to identify a q13 region of a human chromosome 15 and a central region of a mouse chromosome 2.